Analysis of the NF-κB p50 dimer interface by diversity screening

Hart, Darren J., Speight, Robert E., Sutherland, John D. and Blackburn, Jonathan M. (2001) Analysis of the NF-κB p50 dimer interface by diversity screening. Journal of Molecular Biology, 310 3: 563-575. doi:10.1006/jmbi.2001.4724


Author Hart, Darren J.
Speight, Robert E.
Sutherland, John D.
Blackburn, Jonathan M.
Title Analysis of the NF-κB p50 dimer interface by diversity screening
Journal name Journal of Molecular Biology   Check publisher's open access policy
ISSN 0022-2836
1089-8638
Publication date 2001-07-13
Sub-type Article (original research)
DOI 10.1006/jmbi.2001.4724
Volume 310
Issue 3
Start page 563
End page 575
Total pages 13
Place of publication London, United Kingdom
Publisher Academic Press
Language eng
Formatted abstract
An in vivo screen has been devised for NF-κB p50 activity in Escherichia coli exploiting the ability of the mammalian transcription factor to emulate a prokaryotic repressor. Active intracellular p50 was shown to repress the expression of a green fluorescent protein reporter gene allowing for visual screening of colonies expressing active p50 on agar plates. A library of mutants was constructed in which the residues Y267, L269, A308 and V310 of the dimer interface were simultaneously randomised and twenty-five novel functional interfaces were selected which repressed the reporter gene to similar levels as the wild-type protein. The leucine-269 alanine-308 core was repeatedly, but not exclusively, selected from the library whilst a diversity of predominantly non-polar residues were selected at positions 267 and 310. These results indicate that L269 and A308 may form a hot spot of interaction and allow an insight into the processes of dimer selectivity and evolution within this family of transcription factors.
Keyword NF-kB
Homodimer
Random mutagenesis
Dimer interface
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Australian Institute for Bioengineering and Nanotechnology Publications
 
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Created: Wed, 11 Sep 2013, 14:42:51 EST by Cathy Fouhy on behalf of Aust Institute for Bioengineering & Nanotechnology